LOCUS       sequence-237393-        4665 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pMAGIC R4-R3 entry plasmid, contains 2x NLS/p300core scaffold 
            enabling addition of dCas9 mutants into pMAGIC. dCas9 can be 
            inserted via unique SrfI/PmeI restriction sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4665)
  AUTHORS   Haldeman JM, Conway AE, Arlotto ME, Slentz DH, Muoio DM, Becker TC, 
            Newgard CB
  TITLE     Creation of versatile cloning platforms for transgene expression and
            dCas9-based epigenome editing.
  JOURNAL   Nucleic Acids Res. 2018 Dec 27. pii: 5264291. doi: 
            10.1093/nar/gky1286.
  PUBMED    30590691
REFERENCE   2  (bases 1 to 4665)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res. 2018 Dec 27. pii: 5264291. doi: 10.1093/nar/gky1286."
COMMENT     Sequence Label: KL301: pMAGIC (R4-R3) NLS-(SrfI/PmeI)-NLS-p300...
FEATURES             Location/Qualifiers
     source          1..4665
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(603..726)
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      726..735
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             738..758
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             813..860
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             873..2723
                     /codon_start=1
                     /gene="EP300"
                     /product="catalytic histone acetyltransferase core of the 
                     human E1A-associated protein p300 (Delvecchio et al., 
                     2013)"
                     /label=p300 core
                     /translation="IFKPEELRQALMPTLEALYRQDPESLPFRQPVDPQLLGIPDYFDI
                     VKSPMDLSTIKRKLDTGQYQEPWQYVDDIWLMFNNAWLYNRKTSRVYKYCSKLSEVFEQ
                     EIDPVMQSLGYCCGRKLEFSPQTLCCYGKQLCTIPRDATYYSYQNRYHFCEKCFNEIQG
                     ESVSLGDDPSQPQTTINKEQFSKRKNDTLDPELFVECTECGRKMHQICVLHHEIIWPAG
                     FVCDGCLKKSARTRKENKFSAKRLPSTRLGTFLENRVNDFLRRQNHPESGEVTVRVVHA
                     SDKTVEVKPGMKARFVDSGEMAESFPYRTKALFAFEEIDGVDLCFFGMHVQEYGSDCPP
                     PNQRRVYISYLDSVHFFRPKCLRTAVYHEILIGYLEYVKKLGYTTGHIWACPPSEGDDY
                     IFHCHPPDQKIPKPKRLQEWYKKMLDKAVSERIVHDYKDIFKQATEDRLTSAKELPYFE
                     GDFWPNVLEESIKELEQEEEERKREENTSNESTDVTKGDSKNAKKKNNKKTSKNKSSLS
                     RGNKKKPGMPNVSNDLSQKLYATMEKHKEVFFVIRLIAGPAANSLPPIVDPDPLIPCDL
                     MDGRDAFLTLARDKHLEFSSLRRAQWSTMCMLVELHTQSQD"
     regulatory      2487..2496
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2728..2851
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2906..2925)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2907..2925)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2930..2946)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2930..2946)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3048..3067)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3059..3868
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(3126..3145)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4015..4603
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4504..4523
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccct acaggtcact aataccatct aagtagttga
      601 ttcatagtga ctggatatgt tgtgttttac agtattatgt agtctgtttt ttatgcaaaa
      661 tctaatttaa tatattgata tttatatcat tttacgtttc tcgttcaact tttctataca
      721 aagttgccac catggcccca aagaagaagc ggaaggtcgg tatccacgga gtcccagcag
      781 cccgggcgag accatgctcg cggtctcagt ttaaacggcc ggcagccacg aagaaggccg
      841 gccaggcaaa aaagaaaaag ggaggcggga gcattttcaa accagaagaa ctacgacagg
      901 cactgatgcc aactttggag gcactttacc gtcaggatcc agaatccctt ccctttcgtc
      961 aacctgtgga ccctcagctt ttaggaatcc ctgattactt tgatattgtg aagagcccca
     1021 tggatctttc taccattaag aggaagttag acactggaca gtatcaggag ccctggcagt
     1081 atgtcgatga tatttggctt atgttcaata atgcctggtt atataaccgg aaaacatcac
     1141 gggtatacaa atactgctcc aagctctctg aggtctttga acaagaaatt gacccagtga
     1201 tgcaaagcct tggatactgt tgtggcagaa agttggagtt ctctccacag acactgtgtt
     1261 gctacggcaa acagttgtgc acaatacctc gtgatgccac ttattacagt taccagaaca
     1321 ggtatcattt ctgtgagaag tgtttcaatg agatccaagg ggagagcgtt tctttggggg
     1381 atgacccttc ccagcctcaa actacaataa ataaagaaca attttccaag agaaaaaatg
     1441 acacactgga tcctgaactg tttgttgaat gtacagagtg cggaagaaag atgcatcaga
     1501 tctgtgtcct tcaccatgag atcatctggc ctgctggatt cgtctgtgat ggctgtttaa
     1561 agaaaagtgc acgaactagg aaagaaaata agttttctgc taaaaggttg ccatctacca
     1621 gacttggcac ctttctagag aatcgtgtga atgactttct gaggcgacag aatcaccctg
     1681 agtcaggaga ggtcactgtt agagtagttc atgcttctga caaaaccgtg gaagtaaaac
     1741 caggcatgaa agcaaggttt gtggacagtg gagagatggc agaatccttt ccataccgaa
     1801 ccaaagccct ctttgccttt gaagaaattg atggtgttga cctgtgcttc tttggcatgc
     1861 atgttcaaga gtatggctct gactgccctc cacccaacca gaggagagta tacatatctt
     1921 acctcgatag tgttcatttc ttccgtccta aatgcttgag gactgcagtc tatcatgaaa
     1981 tcctaattgg atatttagaa tatgtcaaga aattaggtta cacaacaggg catatttggg
     2041 catgtccacc aagtgaggga gatgattata tcttccattg ccatcctcct gaccagaaga
     2101 tacccaagcc caagcgactg caggaatggt acaaaaaaat gcttgacaag gctgtatcag
     2161 agcgtattgt ccatgactac aaggatattt ttaaacaagc tactgaagat agattaacaa
     2221 gtgcaaagga attgccttat ttcgagggtg atttctggcc caatgttctg gaagaaagca
     2281 ttaaggaact ggaacaggag gaagaagaga gaaaacgaga ggaaaacacc agcaatgaaa
     2341 gcacagatgt gaccaaggga gacagcaaaa atgctaaaaa gaagaataat aagaaaacca
     2401 gcaaaaataa gagcagcctg agtaggggca acaagaagaa acccgggatg cccaatgtat
     2461 ctaacgacct ctcacagaaa ctatatgcca ccatggagaa gcataaagag gtcttctttg
     2521 tgatccgcct cattgctggc cctgctgcca actccctgcc tcccattgtt gatcctgatc
     2581 ctctcatccc ctgcgatctg atggatggtc gggatgcgtt tctcacgctg gcaagggaca
     2641 agcacctgga gttctcttca ctccgaagag cccagtggtc caccatgtgc atgctggtgg
     2701 agctgcacac gcagagccag gacggcgcaa ctttgtataa taaagttgaa cgagaaacgt
     2761 aaaatgatat aaatatcaat atattaaatt agattttgca taaaaaacag actacataat
     2821 actgtaaaac acaacatatc cagtcactat gaatcaacta cttagatggt attagtgacc
     2881 tgtactgcag ggcggccgcg atatccccta tagtgagtcg tattacatgg tcatagctgt
     2941 ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt tacattgcac aagataaaaa
     3001 tatatcatca tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat
     3061 gagccatatt caacgggaaa cgtcgaggcc gcgattaaat tccaacatgg atgctgattt
     3121 atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt
     3181 gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa
     3241 tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac
     3301 catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatccccgg
     3361 aaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc
     3421 gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag
     3481 cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc
     3541 gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca
     3601 taaacttttg ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa
     3661 ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc
     3721 agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt
     3781 acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt
     3841 tcatttgatg ctcgatgagt ttttctaatc agaattggtt aattggttgt aacactggca
     3901 gagcattacg ctgacttgac gggacggcgc aagctcatga ccaaaatccc ttaacgtgag
     3961 ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     4021 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     4081 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     4141 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     4201 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     4261 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     4321 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     4381 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     4441 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     4501 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     4561 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     4621 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgtt
//
